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Abstract

Bottom-up synthetic biology has set itself the exciting and ambitious task to create cellular life from scratch. In particular, the successful mimicry of cell division, which is widely considered one of its fundamental features, represents a milestone towards this goal. However, the complexity of cellular division machineries limits the progress of their reconstitution into synthetic cells, opening the way for novel and creative approaches for the bottom-up engineering of synthetic cell division. \\ In this thesis I engineer mechanisms which can be employed for the implementation and analysis of a synthetic self-replicating cell. In particular, I develop mechanisms for the division of lipid vesicles as synthetic cell compartments with full spatio-temporal control. I present a module which allows for the segregation of DNA droplets as information carriers within synthetic cell compartments. Furthermore, I developed a DNA origami-based genotype-phenotype system which holds important features of natural genotype-phenotpye maps, namely, variability and stability. Additionally, I developed an open-source software tool for 3D object reconstruction and analysis to study structural changes during division and phenotypic effects on vesicle morphology.\\ All in all, these contributions provide valuable strategies for the implementation and analysis of a synthetic self-replicating cell, bringing us closer to the ultimate goal of creating life from scratch.